The continuing goal of this project is to produce reliable and large-scale supplies of medical grade, quality-controlled Keyhole Limpet Hemocyanin (KLH) products ready for use in an expanding array of cancer vaccines, immunological uses and other biomedical applications supported by the NIH and private industry. With the assistance of funds from Phase l and Phase II SBIR Grants from the National Center for Research Resources, Stellar Biotechnologies has successfully developed, optimized and demonstrated the first dedicated aquaculture technology and non-lethal hemocyanin extraction methods for commercial scale production-of KLH from Megathura crenulata (the giant keyhole limpet) for biomedical applications. We have met and exceeded quality standards and production schedule requirements of two manufacturers of cancer vaccines in clinical trials. Market research conducted during our Phase II SBIR research identified therapeutic cancer vaccines as the leading and most tractable application for Stellar's KLH products, and identified specific requirements that will be necessary for a commercially viable scale of production. To meet these requirements and make cGMP (current Good Manufacturing Practices) compliant KLH reliably available to support the burgeoning applications for cancer vaccines, we propose here to complete the commercial-scale development of production processes, facilities, and quality systems necessary to refine Stellar's KLH products and manufacturing capabilities to meet the specifications and volume requirements of therapeutic cancer vaccine developers and the regulatory standards of the FDA. We propose three specific aims in order to accomplish these goals: 1) to develop standardized, high-purity, quality controlled monomeric and defined multimeric KLH formulations supported by a large-scale and reliable supply of hemolymph from the company's proprietary high-yield aquaculture system; 2) to renovate our existing facilities to include a Clean Room to house production operations for high-purity medical grade KLH in compliance with FDA's cGMP regulations for the manufacture of such products for cancer vaccines, other pharmaceuticals and in vitro diagnostics; and 3) to submit a Type II Drug Master File to the FDA in order to facilitate the use of KLH by prospective manufacturers of cancer vaccines and other medical products. The outcome of this project will be a domestic aquaculture-based cGMP KLH production facility with the capability of supplying highly characterized and price-stabilized commercial quantities of the two forms of this critical immunostimulatory molecule most required by cancer vaccine developers. This project directly addresses and alleviates the present supply risks associated with the use of KLH produced from the limited population of keyhole limpets harvested from the wild, and will thus facilitate medical use and further biomedical explorations of this important immunostimulatory molecule. [unreadable] [unreadable] [unreadable]